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APC Rat anti-Mouse CD40
APC Rat anti-Mouse CD40
Flow cytometric analysis of APC-conjugated anti-mouse CD40 on mouse splenocytes. Murine splenocytes were stained with FITC conjugated anti-mouse CD4/CD8a (clones RM4-5 /53-6.7 respectively; Cat. No. 553046/553030) and either APC anti-CD40 (clone 3/23, Cat. No. 558695, right panel) or an APC rat IgG2a isotype control (Cat. No. 553932, left panel) and analyzed by flow cytometry.  Flow cytometry was performed on a BD FACSCalibur™ System and the dot plots were derived from the gated events based on light scattering characteristics of viable splenocytes.
Flow cytometric analysis of APC-conjugated anti-mouse CD40 on mouse splenocytes. Murine splenocytes were stained with FITC conjugated anti-mouse CD4/CD8a (clones RM4-5 /53-6.7 respectively; Cat. No. 553046/553030) and either APC anti-CD40 (clone 3/23, Cat. No. 558695, right panel) or an APC rat IgG2a isotype control (Cat. No. 553932, left panel) and analyzed by flow cytometry.  Flow cytometry was performed on a BD FACSCalibur™ System and the dot plots were derived from the gated events based on light scattering characteristics of viable splenocytes.
Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat LOU, also known as Louvain, LOU/C, LOU/M IgG2a, κ
Mouse CD40 Recombinant Protein
Flow cytometry (Routinely Tested)
0.2 mg/ml
21939
AB_1645224
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The polyclonal antibody was purified from antiserum by affinity chromatography. The antibody was conjugated to APC under optimum conditions, and unconjugated antibody and free APC were removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
558695 Rev. 1
Antibody Details
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3/23

The 3/23 clone monoclonal antibody specifically binds to CD40, a 40-50 kDa glycoprotein expressed on B lymphocytes and other antigen-presenting cells.  CD40 has been reported to be transiently expressed on activated CD4+ and CD8+ T cells and in some mouse strains, the 3/23 mAb has been reported to react with 5-10% of T lymphocytes in adult mouse, but not neonatal, spleen.  CD40 plays a key role in B-cell growth and differentiation where interactions of CD40 with its ligand, CD154, are involved in the initiation, effector, and memory stages of cell-mediated immune responses.  In addition, CD40 has been reported to be involved with the triggering of NK cells and NK-T cells. Ligation of CD40 with the 3/23 antibody has been reported to induce splenic B cells to express the costimulatory molecule CD86 (B7-2).  In addition, although the 3/23 antibody by itself is a weak B-cell mitogen, it has been reported to synergize markedly with mitogenic anti-IgM, anti-IgD mAb or IL-4 to promote B-cell proliferation.

558695 Rev. 1
Format Details
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APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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APC
Red 627-640 nm
651 nm
660 nm
558695 Rev.1
Citations & References
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Development References (13)

  1. Bourgeois C, Rocha B, Tanchot C. A role for CD40 expression on CD8+ T cells in the generation of CD8+ T cell memory. Science. 2002; 297(5589):2060-2063. (Biology). View Reference
  2. French RR, Chan HT, Tutt AL, Glennie MJ. CD40 antibody evokes a cytotoxic T-cell response that eradicates lymphoma and bypasses T-cell help. Nat Med. 1999; 5(5):548-553. (Clone-specific). View Reference
  3. Grewal IS, Flavell RA. CD40 and CD154 in cell-mediated immunity. Annu Rev Immunol. 1998; 16:111-135. (Biology). View Reference
  4. Hasbold J, Johnson-Leger C, Atkins CJ, Clark EA, Klaus GG. Properties of mouse CD40: cellular distribution of CD40 and B cell activation by monoclonal anti-mouse CD40 antibodies. Eur J Immunol. 1994; 24(8):1835-1842. (Immunogen: Activation, (Co)-stimulation). View Reference
  5. Hasbold J, Klaus GG. B cells from CBA/N mice do not proliferate following ligation of CD40. Eur J Immunol. 1994; 24(1):152-157. (Biology: Activation, (Co)-stimulation). View Reference
  6. Klaus GG, Holman M, Hasbold J. Properties of mouse CD40: the role of homotypic adhesion in the activation of B cells via CD40. Eur J Immunol. 1994; 24(11):2714-2719. (Biology). View Reference
  7. Leifeld L, Trautwein C, Dumoulin FL, Manns MP, Sauerbruch T, Spengler U. Enhanced expression of CD80 (B7-1), CD86 (B7-2), and CD40 and their ligands CD28 and CD154 in fulminant hepatic failure. Am J Pathol. 1999; 154(6):1711-1720. (Biology: Immunohistochemistry). View Reference
  8. Mahnke K, Becher P, Ricciardi-Castagnoli P, Luger TA, Schawrz T Grabbe S. CD14 is expressed by subsets of murine dendritic cells and upregulated by lipopolysaccharide. In: Ricciardi-Castagnoli P, ed. Dendritic Cells in Fundamental and Clinical Immunology. New York: Plenum Press; 1997:145-159.
  9. Munder M, Mallo M, Eichmann K, Modolell M. Murine macrophages secrete interferon gamma upon combined stimulation with interleukin (IL)-12 and IL-18: A novel pathway of autocrine macrophage activation. J Exp Med. 1998; 187(12):2103-2108. (Biology). View Reference
  10. Noelle RJ, Ledbetter JA, Aruffo A. CD40 and its ligand, an essential ligand-receptor pair for thymus-dependent B-cell activation. Immunol Today. 1992; 13(11):431-433. (Biology). View Reference
  11. Parry SL, Hasbold J, Holman M, Klaus GG. Hypercross-linking surface IgM or IgD receptors on mature B cells induces apoptosis that is reversed by costimulation with IL-4 and anti-CD40. J Immunol. 1994; 152(6):2821-2829. (Biology). View Reference
  12. Parry SL, Holman MJ, Hasbold J, Klaus GG. Plastic-immobilized anti-mu or anti-delta antibodies induce apoptosis in mature murine B lymphocytes. Eur J Immunol. 1994; 24(4):974-979. (Biology). View Reference
  13. Tomura M, Yu WG, Ahn HJ, et al. A novel function of Valpha14+CD4+NKT cells: stimulation of IL-12 production by antigen-presenting cells in the innate immune system. J Immunol. 1999; 163(1):93-101. (Biology). View Reference
View All (13) View Less
558695 Rev. 1

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